The outbreak of the coronavirus disease 2019 (COVID-19) pandemic has sparked significant concerns regarding the significance of bioaerosols
. Assessing air quality and identifying microbial pollution is crucial for estimating health risks and establishing air quality control standards. This research aimed to comprehensively investigate the microbial composition of the air at selected sites in Nigeria, focusing on the presence of potential pathogenic organisms. Over a duration of six months, from October 2019 to March 2020, samples were collected using settle plates employing the open-plate method. Standard Petri dishes containing culture media were exposed to the air for 10 to 15 minutes, facilitating the collection of settled biological particles. The designated locations included two interstate travel motor parks and two marketplaces situated in Ede and Osogbo, Osun State, Nigeria. The isolates underwent preliminary identification tests, followed by further identification of bacterial isolates using molecular techniques were employed including DNA amplification of the 16S rDNA gene using polymerase chain reaction (PCR), agarose gel electrophoresis of PCR amplicons, Sanger sequencing of bacterial 16S rDNA gene, and subsequent phylogenetic and evolutionary analysis using MEGA X. The results revealed the presence of diverse microorganisms, including pathogenic species, highlighting the importance of monitoring air quality for public health. This study contributes to the understanding of microbial pollution in the air and provides valuable insights for air quality management strategies.